PA5-26323

antibody from Invitrogen Antibodies

Targeting: GPX1

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-26323

Provider

Invitrogen Antibodies

Product name

GPX1 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

400 µL

Concentration

2.0 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Retinal oxidative stress activates the NRF2/ARE pathway: An early endogenous protective response to ocular hypertension.

Naguib S, Backstrom JR, Gil M, Calkins DJ, Rex TS
Redox biology 2021 Jun;42:101883

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Oxidative stress and mitochondrial uncoupling protein 2 expression in hepatic steatosis induced by exposure to xenobiotic DDE and high fat diet in male Wistar rats.

Migliaccio V, Scudiero R, Sica R, Lionetti L, Putti R
PloS one 2019;14(4):e0215955

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of HepG2 cells using a GPX1 polyclonal antibody (Product # PA5-26323) at a dilution of 1:10-50, followed by a fluor-conjugated goat anti-rabbit secondary antibody (green). Nuclei were stained with DAPI (blue).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of HeLa cells using a GPX1 polyclonal antibody (Product # PA5-26323) (bottom) compared to a negative control cell (top) at a dilution of 1:10-50, followed by a FITC-conjugated goat anti-rabbit antibody

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 2 ROS and antioxidant proteins increased at 1-wk and 2-wks post-IOP elevation. A) Representative fundus images of DHE fluorescence in all groups. B) Quantification of DHE fluorescence at 1-4 wks post-IOP elevation in comparison to saline-injected controls, showing increases in the microbead injected mice at all time points, **p < 0.001, ****p < 0.0001. C) Fluorescence micrographs of retinas labeled with DAPI (blue) and nitrotyrosine (green). Inset shows double-labeling with anti-nitrotyrosine (green) and anti-NeuN (red). D) Quantification of nitrotyrosine immunofluorescence showing increases at all timepoints after IOP elevation, ***p < 0.001, ****p < 0.0001. E-G) Quantification of fold changes in PCR microarray for: E) GSH-related genes; F) Peroxiredoxin-related genes; and G) SOD-related genes. Dotted line indicates threshold for increase compared to saline. H) Confirmatory qPCR for fold change over saline in antioxidant gene transcription, *p < 0.05, **p < 0.01. I) Representative western blots for beta-actin, PRDX6, SOD3 and GPX1. J-L) Quantification of PRDX6, SOD3 and GPX1, respectively, after normalization to beta-actin, *p < 0.05, **p < 0.001. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.) Fig. 2

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 5 PI3K-dependent NRF2 phosphorylation. A) Representative western blots of PI3K and pPI3K B) Quantification of pPI3K to total PI3K, ****p < 0.00001. C) Experiment timeline. D) Representative western blots of pNRF2 to total NRF2. E) Quantification of pNRF2 to total NRF2, ****p < 0.00001. F) Quantification of antioxidant gene transcription shown as fold change over saline, *p < 0.05, ***p < 0.0001. G) Representative western blots for beta-actin, PRDX6, SOD3 and GPX1. H-J) Quantification of PRDX6, GPX1 and SOD3, respectively, after normalization to beta-actin, *p < 0.05, ***p < 0.0001. Fig. 5

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 6 Decreased antioxidants and increased ROS, axon degeneration, and visual function deficits in ocular hypertensive Nrf2 KO mice. A) Representative fundus images of DHE fluorescence in saline and microbead-injected wildtype and Nrf2 KO mice. B) Quantification of DHE fluorescence at 2-wks post-IOP elevation in both strains of mice. C) Quantification of antioxidant gene transcription shown as fold change over respective saline groups, *p < 0.05, **p < 0.001, ***p < 0.0001. D) Representative western blots for beta-actin, PRDX6, SOD3 and GPX1. E-G) Quantification of PRDX6, GPX1 and SOD3, respectively, after normalization to beta-actin, **p < 0.001. H) Representative micrographs of wildtype and Nrf2 KO optic nerves at 2-wks post-IOP elevation. Scale bar applies to all micrographs. Arrows indicate degenerative axons. I) Quantification of total axons in the optic nerves, showing a decrease only in ocular hypertensive Nrf2 KO mice, **p < 0.001. J) Quantification of number of degenerative axons, showing an increase in ocular hypertensive Nrf2 KO mice, ***p < 0.0001. K) Quantification of the N1 amplitude of the VEP. L) Quantification of the PhNR amplitude. Note: all wild-type data shown in this Figure was copied from Fig. 2 for purposes of comparison. Fig. 6